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An Easy Guide for Practical Biochemistry
Divya Shanthi D’sa, Sowbhagya Lakshmi
SECTION 1: LABORATORY RULES AND REGULATIONS
1:
Laboratory Hazards and First Aid
HAZARDS
SAFETY MEASURES
CARE WHILE PIPETTING
PRECAUTIONS REGARDING FIRE SAFETY
IN CASE OF ACCIDENTAL FIRE
ACCIDENTAL SWALLOW OF CORROSIVE SOLUTIONS
INHALATION OF CORROSIVE GASES
BURNS
LABORATORY FIRST AID
PRECAUTIONS TO BE TAKEN WHILE HANDLING CHEMICALS
IN CASE OF ACCIDENTS IN THE LABORATORY
WASTE DISPOSAL
Chemical Waste
Radioactive Waste
2:
Laboratory Safety Rules
GENERAL GUIDELINES
ACCIDENTS AND INJURIES
HANDLING CHEMICALS
HANDLING GLASSWARE AND EQUIPMENT
HEATING SUBSTANCES
AFTER EXPERIMENTS
PIPETTING TECHNIQUES
3:
Specimen Collection and Processing
COLLECTION OF BLOOD
ANTICOAGULANTS
COLLECTION AND PRESERVATION OF URINE SPECIMENS
Collection of Urine Samples
Preservation of Urine Samples
4:
Glasswares Used in Biochemistry Laboratory
LABORATORY GLASSWARES
Beakers
Flasks
Graduated (Measuring) Cylinder
Burettes
Funnels
Bottles
Reagent Bottles
Drop Bottles
Wash Bottles
PIPETTES
Test Tubes
Centrifuge Tubes
Folin-Wu Tubes
Dispensers
Desiccators
CLEANING OF GLASSWARE
SECTION 2: QUALITATIVE TESTS
5:
Qualitative Analysis of Carbohydrates
DEFINITION
CLASSIFICATION
Monosaccharides
Oligosaccharides
Polysaccharides
FUNCTIONS OF CARBOHYDRATES
REACTIONS OF CARBOHYDRATES
Reaction with Alkalies
Weak Alkalies
Strong Alkalies
Reaction with Acids
Weak Acids
Strong Acids
TESTS FOR CARBOHYDRATES
Physical Properties
Chemical Tests
Molisch Test
Iodine Test
Benedict's Test (Reduction under alkaline condition)
Benedict's tests (In case of sucrose)
Acid Hydrolysis
Benedict's test (In case of starch)
Barfoed's Test (Reduction under acidic medium)
Seliwanoff's Test
Seliwanoff's test (for sucrose)
Osazone Test
Importance and significance
6:
Qualitative Analysis of Proteins
DEFINITION
CLASSIFICATION
FUNCTIONS OF PROTEINS
REACTIONS OF PROTEINS
Precipitation Reactions of Proteins
Types of Colloids
Importance of Precipitation of Proteins
Amino Acid
Precipitation by Salts
Isoelectric Precipitation
Precipitation by Organic Solvents
Precipitation by Alkaloidal Reagents
Precipitation by Heavy Metal Ions
Precipitation by Heat and Acid (Heat and Acetic Acid Test)
COLOR REACTIONS FOR PROTEINS
Albumin
Functions of Albumin
Casein
Physical Properties of Albumin
Physical Properties of Casein
Chemical Properties
Biuret Test
Ninhydrin Test
Xanthoproteic Test
Millon's Test (Cole's mercuric nitrite test)
Aldehyde Test for Indole Nucleus (Hopkins-Cole-Adam's Test)
Sakaguchis Test for Guanidine Group
Sulfur Test for Cystine and Cysteine
Pauly's Test for Histidine and Tyrosine
Molisch Test for Carbohydrate Moiety in Proteins
Test for Organic Phosphorus (Neumann's Test) (Test with casein solution)
7:
Nonprotein Nitrogenous Substances
8:
Qualitative Analysis of Normal Urine
EXAMINATION OF URINE
SPECIMEN COLLECTION
PRESERVATION OF URINE SAMPLES
GENERAL AND PHYSICAL CHARACTERISTICS
Volume
Appearance
Color
Odor
Specific Gravity
CHEMICAL CHARACTERISTICS
Reaction
Constituents of Normal Urine
ANALYSIS OF NORMAL URINE
Physical Examination
Determination of Specific Gravity
Long's Coefficient
Chemical Tests
Inorganic Constituents
Test for Chloride
Test for Inorganic Sulfates
Test for Phosphates and Calcium
Test for Ammonia
Organic Constituents
Test for Urea
Test for Uric Acid
Test for Creatinine (Jaffe's Test)
Test for Ethereal Sulfate (Organic Sulfate)
9:
Analysis of Abnormal Constituents in Urine
ANALYSIS OF ABNORMAL URINE
Physical Characteristics
Chemical Constituents
Test for Proteins
Heat and Acetic Acid Test
Heller's Nitric Acid Ring Test
Sulfosalicylic Acid Test
Test for Reducing Sugar (Benedict's Test)
Test for Ketone Bodies
Rothera's Test for Acetone and Acetoacetic Acid
Gerhardt's Test for Acetoacetic Acid
Test for Bile Salts (Hay's Test)
Test for Bile Pigments
Gmelin's Test
Fouchet's Test
Test for Blood
10:
Hemoglobin and its Derivatives
DETECTION OF HEMOGLOBIN AND ITS DERIVATIVES
Direct Vision Spectroscope
Principle
PREPARATION OF HEMOGLOBIN AND DERIVATIVES
Oxyhemoglobin (HbO2)
Reduced Hemoglobin
Carboxy Hemoglobin
Methemoglobin
Points to Note
Preparation of Hemin Crystals
11:
Spot Tests
PHENYLKETONURIA
Guthrie's Bacterial Inhibition Test
Ferric Chloride Test
Alkaptonuria
Spot Test
Procedure
HOMOCYSTINURIA
Screening Test for Homocystinuria (Spaeth and Barber)
Reagents
Procedure
SECTION 3: QUANTITATIVE TESTS
12:
Principles of Colorimetry
PHOTOMETRY
Principle
COLORIMETER
Components of the Colorimeter
PREPARATION OF SOLUTION FOR INVESTIGATION
Blank
Type of Blank
Standard Solution
Test Solution
TECHNIQUE
Beer's Law
Lambert's Law
RELATIONSHIP BETWEEN ABSORBANCE AND TRANSMITTANCE
SELECTION OF FILTER IN COLORIMETRIC ESTIMATION
Steps in the Operation of the Colorimeter
CALCULATIONS
APPLICATION OF COLORIMETER
13:
Estimation of Blood Sugar
14:
Estimation of Blood Urea
15:
Estimation of Urine Creatinine
16:
Estimation of Serum Inorganic Phosphate
17:
Estimation of Serum Total Proteins
SECTION 4: DEMONSTRATION PRACTICALS
18:
Chromatography
19:
Electrophoresis
20:
Glucose Tolerance Test
21:
Estimation of Serum AST and ALT
22:
Estimation of Serum Cholesterol
23:
Estimation of Plasma Ascorbic Acid
24:
Flame Photometer
25:
CSF Analysis
26:
Estimation of Albumin in Urine
Appendices
NECESSITY OF QUALITY CONTROL
SOME IMPORTANT TERMS
Precision
Accuracy
Specificity
Sensitivity
Standard
Control
VARIANCE
STANDARD DEVIATION (SD)
COEFFICIENT OF VARIATION (CV)
QUALITY CONTROL CHART (LEVEY-JENNINGS CHART)
READING THE CHART
PREPARATION OF QUALITY CONTROL (QC) MATERIAL IN THE LABORATORY
QUALITY CONTROL PROGRAMS
INTERNAL AND EXTERNAL QUALITY CONTROL
Internal Quality Control
External Quality Control
TYPES OF ERRORS OBSERVED AND THEIR CORRECTION
Intrinsic Errors
Systemic Errors
Random /Technical Errors
PREANALYTICAL ERRORS
ANALYTICAL ERRORS
POSTANALYTICAL ERRORS
MOLECULAR WEIGHT
GRAM MOLECULAR WEIGHT
MOLALITY
MOLARITY
EQUIVALENT WEIGHT
NORMALITY
SOLUTIONS
Standard Solution
Normal Solution
Molar Solution
Percent Solution
Saturated Solution
Reagent Solution
Stock Reagent
PREPARATION OF NORMAL SOLUTIONS
EQUIVALENT WEIGHT OF AN ACID
CONVERSION CHART
INTERNAL ASSESSMENT FOR BIOCHEMISTRY
THEORY AND RECORDS
PRACTICALS
UNIVERSITY EXAMINATION
PRACTICAL EXAMINATION: 40 MARKS
Exercise I: Two hours, 20 marks
Exercise II: Two hours, 20 marks
Viva Voce Examination: 20 Marks
INDEX
TOC
Index
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